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Primers for detecting methylation degree of BDNF gene promoter region and application of primers

A methylation and promoter technology, applied in the fields of biotechnology and neurocognition, can solve the problems of undiscovered BDNF gene, etc., and achieve the effects of convenient and fast detection, improved sensitivity and specificity, and accurate quantification

Inactive Publication Date: 2017-03-29
THE FIRST HOSPITAL OF HEBEI MEDICAL UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, the current research has not found that BDNF gene is specifically related to AD.

Method used

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  • Primers for detecting methylation degree of BDNF gene promoter region and application of primers
  • Primers for detecting methylation degree of BDNF gene promoter region and application of primers
  • Primers for detecting methylation degree of BDNF gene promoter region and application of primers

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0043] Example 1 Establishing a method for predicting the outcome of mild cognitive impairment to Alzheimer's disease

[0044] This application provides a method for establishing and predicting the outcome of mild cognitive impairment to Alzheimer's disease, which is carried out according to the following steps:

[0045] (1) Establishment of demographic database, clinical database and clinical specimen database

[0046] Baseline and follow-up demographic data, clinical data, and blood samples were collected using standard operating procedures:

[0047] Baseline collection: including collection of demographic data, clinical data and blood samples from normal cognitive subjects and aMCI patients over 60 years old in the mild cognitive impairment baseline survey database;

[0048] Collection of follow-up research subjects: In the baseline survey database of mild cognitive impairment, demographic data, clinical data and blood samples were collected from aMCI patients over 60 year...

Embodiment 2

[0054] Example 2 Selection of the BDNF gene promoter region

[0055] Find And determine the promoter region of BDNF gene. Determine the promoter region as promoter I and promoter IV.

[0056] The sequence of promoter I is as follows, the gray shading part is the site to be tested, in which: the three CG sequences from left to right in this part are CpG1-CpG3 respectively:

[0057]

[0058] The sequence of promoter IV is as follows, the gray shading is the site to be tested, where: the 4 CG sequences from left to right in this part are CpG4-CpG7 respectively:

[0059]

Embodiment 3

[0060] The design of embodiment 3 methylation-specific PCR primers and pyrosequencing primers

[0061] Using the Meth Primer primer design software (http: / / www.urogene.org / methprimer / index1.html), design the corresponding methylation-specific primers and pyrosequencing primers for the target region to be detected according to the experimental requirements, and design Good methylation-specific primers were further verified by methBLAST (http: / / medgen.ugent.be / methBLAST / ) to ensure the specificity of the target amplified sequence. Primers were designed as follows:

[0062] Promoter IV primer

[0063]

[0064] Promoter I primer

[0065]

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Abstract

The invention discloses primers for detecting the methylation degree of a BDNF gene promoter region and application of the primers. The primers comprise the BDNF gene promoter region methylation specific amplification primer and the pyrosequencing primer, the methylation specific upstream primer nucleotide sequence is shown in SEQ ID NO.1, the reverse primer has the sequence shown in SEQ ID NO.2, and the pyrosequencing primer sequence is shown in SEQ ID NO.3. The primers have the advantage that a prediction kit can stably, noninvasively, conveniently and quickly predict lapse from mild cognitive impairment to the Alzheimer disease at the molecular level in accurate-quantification, high-sensitivity and good-specificity modes.

Description

technical field [0001] The invention relates to the field of biotechnology and neurocognitive technology, in particular to a kit for preparing primers required for detecting the degree of methylation of the BDNF gene promoter region and predicting the progression from mild cognitive impairment to Alzheimer's disease in the application. Background technique [0002] Alzheimer's disease (AD) is a neurodegenerative disease that seriously threatens human health. Mild cognitive impairment (MCI) is a transitional state between AD and normal aging. According to clinical manifestations, MCI is divided into amnestic and non-amnestic types, among which amnestic MCI (amnestic MCI, aMCI) is the most common. Studies have shown that 23% of aMCI patients progress to AD within 2 years, and 50% of patients progress to AD within 4 years, so aMCI is considered to be the most appropriate stage for early intervention and treatment of AD. With the aging of our population, the prevalence of AD ...

Claims

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Application Information

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IPC IPC(8): C12Q1/68C12N15/11
CPCC12Q1/6883C12Q2600/112C12Q2600/154
Inventor 谢冰许顺江张庆富徐垚张睿
Owner THE FIRST HOSPITAL OF HEBEI MEDICAL UNIV
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